Dihydro-isoquinoline compounds and their use as pharmaceuticals

ABSTRACT

Compound of general formula I   &lt;IMAGE&gt; I  wherein A is a benzo or thieno group; R1 is (C4-6)cycloalkyl, (C4-6)cycloalkyl-(C1-5)alkyl or &lt;IMAGE&gt;  R2, m, R3, R4, R and u are defined as in the specification, and pharmaceutical preparations containing this compound and the new pharmaceutical uses thereof.

This is a continuation, of application Ser. No. 08/478,298, filed Jun. 6, 1995 abandoned, which is a divisional of application Ser. No. 08/249,822, filed May 26, 1994 abandoned which is a continuation of application Ser. No. 08/081,599, filed Jun. 22, 1993, abandoned.

The invention relates to anellated dihydropyridinoacetic acid derivatives, processes for preparing them and pharmaceutical compositions containing these compounds.

Dihydroisoquinolines are known from EP-A 37 934. The compounds mentioned therein have a cardiotonic activity and have an activity component which increases contractility and influences blood pressure. They have been proposed for improving circulation of the blood through the tissues and for improving the supply of oxygen to the tissues. These possible uses are based on the vascular activity of the compounds. EP-A 251 194 describes how carbocyclically and heterocyclically anellated dihydropyridines have a cardioprotective activity and constitute an entirely new type of Ca-antagonistic compounds.

The present invention relates to new carbocyclically and heterocyclically anellated dihydropyridines, and the pharmaceutical use of these compounds and new pharmaceutical uses for the dihydroisoquinolines known from EP-A-37 934. These new uses are based on their antiproliferative effect and their activity in the treatment of ulcerative colitis and Crohn's disease.

One aspect of the invention consists in the use of a compound of general formula I ##STR3## wherein A denotes a benzo, indolo or thieno group; wherein, if A is benzo, m is 2 or 3 (preferably 2, the two R² groups being in positions 6 and 7), and the substituents R² independently of each other may represent hydroxy, (C₁₋₄)alkoxy, benzyloxy, halogen (F, Cl, Br, I), (C₁₋₄)alkyl, methanesulphonyloxy or methanesulphonamido, or two adjacent substituents R² together represent --O--CH₂ --O-- or --O--CH₂ --CH₂ --O--; and if A is indolo or thieno, m is zero;

R₁ denotes (C₄₋₆)cycloalkyl, (C₄₋₆)cycloalkyl(C₁₋₅)-alkyl or ##STR4## R³ and R⁴ independently of each other denote (a) hydrogen,

(b) branched or unbranched C₃₋₆ -alkenyl,

(c) branched or unbranched C₃₋₆ -alkynyl or

(d) branched or unbranched C₁₋₁₂ -alkyl, whilst the

alkyl may be substituted by

hydroxy,

(C₁₋₄)alkoxy,

di (C₁₋₄)alkylamino,

furyl,

pyridyl,

pyrrolidinyl, N-methylpyrrolidinyl,

morpholino,

indolyl,

nitrilo,

thienyl,

adamantyl,

cyclohexyl,

phenoxy,

naphthyloxy or phenyl, whilst this phenyl or the phenyl contained in the phenoxy group may be mono-, di- or trisubstituted by hydroxy, (C₁₋₄)alkoxy, benzyloxy, halogen (F, Cl, Br, I), CF₃, N₃, (C₁₋₄)alkyl, adamantyl, --SO₂ NH₂, --NHCOCH₃, --NHSO₂ CH₃ or CH₃ SO₂ O-- or by the bridge --O--CH₂ --O--;

or R³ denotes hydrogen and R⁴ denotes cyclohexyl, phenyl, fluorophenyl, pyridyl or N-benzylpiperidyl;

or R³ and R⁴ together with the nitrogen atom to which they are bound represent pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, the group ##STR5## or piperazinyl, whilst the piperazinyl ring may optionally be N-substituted by methyl, unsubstituted phenyl, mono- or di(C₁₋₄)alkoxyphenyl, pyrimidinyl, phenyl(C₁₋₄)alkyl or ##STR6## R denotes C₁₋₄ -alkyl, hydroxy, --N₃, halogen (F, Cl, Br, I), CF₃, C₁₋₄ -alkoxy or --COH and

u denotes 0, 1, 2 or 3;

A further aspect of the invention consists in the following new compounds:

of general formula ##STR7## wherein R¹, R³ and R⁴ are as defined above; and new compounds of general formula I ##STR8## wherein A denotes a benzo, indolo or thieno group; wherein, if A is benzo, m is 2 or 3 (preferably 2, the two R² groups being in positions 6 and 7), and the substituents R² independently of each other may represent hydroxy, (C₁₋₄)alkoxy, benzyloxy, halogen (F, Cl, Br, I), (C₁₋₄)alkyl, methanesulphonyloxy or methanesulphonamido, or two adjacent substituents R² together represent --O--CH₂ --O or --O--CH₂ --CH₂ --O--; and if A is indolo or thieno, m is zero; if A is indolo or thieno, m is zero;

R₁ denotes (C₄₋₆)cycloalkyl, (C₄₋₆)cycloalkyl(C₁₋₅)-alkyl or ##STR9## R³ and R⁴ independently of each other denote (a) hydrogen,

(b) branched or unbranched C₃₋₆ -alkenyl,

(c) branched or unbranched C₃₋₆ -alkynyl or

(d) branched or unbranched C₁₋₁₂ -alkyl, whilst the

alkyl may be substituted by

hydroxy,

(C₁₋₄) alkoxy,

di (C₁₋₄)alkylamino,

furyl,

pyridyl,

pyrrolidinyl, N-methylpyrrolidinyl,

morpholino,

indolyl,

nitrilo,

thienyl,

adamantyl,

cyclohexyl,

phenoxy,

naphthyloxy or phenyl, whilst this phenyl or the phenyl contained in the phenoxy group may be mono-, di- or trisubstituted by hydroxy, (C₁₋₄)alkoxy, benzyloxy, halogen (F, Cl, Br, I), CF₃, N₃, (C₁₋₄)alkyl, adamantyl, --SO₂ NH₂, --NHCOCH₃, --NHSO₂ CH₃ or CH₃ SO₂ O-- or by the bridge --O--CH₂ --O--;

or R³ denotes hydrogen and R⁴ denotes cyclohexyl, phenyl, fluorophenyl, pyridyl or or R³ and R⁴ together with the nitrogen atom to which they are bound represent

pyrrolidinyl,

piperidinyl,

morpholinyl, thiomorpholinyl, the group ##STR10## or piperazinyl, whilst the piperazinyl ring may optionally be N-substituted by methyl, unsubstituted phenyl, mono- or di(C₁₋₄)alkoxyphenyl, pyrimidinyl, phenyl(C₁₋₄)alkyl or ##STR11## R denotes C₁₋₄ -alkyl, hydroxy, --N₃, halogen (F, Cl, Br, I), CF₃, C₁₋₄ -alkoxy or --COH and

u denotes 0, 1, 2 or 3.

Another aspect of the invention consists in individual new compounds which come under the general definition of earlier patent applications. These compounds are contained, together with other compounds, in Tables 13 to 20, particularly Tables 14, 16 and 20. The following compounds of this group should be particularly mentioned:

New compounds of general formula I of the formula ##STR12## wherein NR³ R⁴ has one of the following meanings

    ______________________________________                              Structure     ______________________________________      ##STR13##                 I      ##STR14##                 I      ##STR15##                 I      ##STR16##                 I + II      ##STR17##                 I      ##STR18##                 I      ##STR19##      ##STR20##     HNCH.sub.2 -1-Adam.      ##STR21##      ##STR22##      ##STR23##      ##STR24##      ##STR25##      ##STR26##                 I      ##STR27##                 I      ##STR28##                 I      ##STR29##                 I      ##STR30##                 I      ##STR31##                 I      ##STR32##                 II      ##STR33##                 I     NHCH.sub.2 -Adam(1)        II      ##STR34##                 I      ##STR35##      ##STR36##     ______________________________________

Compounds of formula I form tautomers of formula II ##STR37##

The tautomers can be separated by known methods, e.g. by column chromatography or selective reduction (NaBH₄ or catalytic reduction).

The compounds of formula II may be present in cis- and/or trans-form: ##STR38##

The invention also comprises the physiologically acceptable salts of the above new compounds with acids, bases and complexing agents.

The new compounds have valuable therapeutically useful properties. They can be used as cardioprotective agents, as cerebroprotective agents (particularly for the treatment of patients who have suffered a stroke or are in danger of suffering a stroke), and as agents for treating chronic inflammatory processes (e.g. bronchial asthma and arthritis). Furthermore, these compounds can be used as agents with an antiproliferative effect and as agents for treating ulcerative colitis and Crohn's disease.

In the definitions used in the text, the radicals and groups may be identical or different, i.e. if one of the above-mentioned substituents occurs several times in one particular molecule, the meaning can be freely selected on each occasion within the scope of the range of definitions.

The term alkyl refers particularly to C₁₋₆ -alkyl and C₁₋₄ -alkyl radicals which may in turn be substituted or, as alkyl radicals, are part of a functional group such as alkoxy or alkylthio. The alkyl radicals include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec.butyl, isobutyl, tert.butyl radicals and the various isomeric pentyl and hexyl radicals such as isopentyl, neopentyl, n-pentyl and the n-hexyl radical.

The above definition thus applies even when the alkyl radical is in turn substituted and/or is itself part of an alkoxyalkyl, alkoxycarbonyl, alkoxy, alkylthio, alkylsulphonyl, monoalkylamino, alkylmethyl, alkylthiomethyl, dialkylamino group or the alkyl radical is bound as a substituent to an aromatic heterocyclic or carbocyclic system.

The halogens are fluorine, chlorine, bromine and iodine, preferably fluorine, chlorine and bromine and, to a lesser extent, iodine.

C₃₋₆ -cycloalkyl means cyclopropane, cyclobutane, cyclopentane and cyclohexane.

C₅₋₆ -cycloalkenes are for example cyclopentene, cyclohexene and cyclohexadiene.

The C₂ - and C₃ -acyl radicals represent the acetyl and propionyl radicals.

C₃₋₆ -alkynes are the isomeric hexynes, pentynes, butynes and propynes, preferably propargyl.

The C₃₋₆ -alkenes are the isomeric hexenes, pentenes, butenes and propenes, preferably allyl.

Examples of unsaturated heterocycles include, inter alia:

furan, pyran, pyrrole, pyrazole, imidazole, pyridine, pyrazine, pyrimidine, pyridazine, thiophene, thiazole, oxazole, 1,2,4-triazole, 1,2,3-triazole, 1,2,4-triazine, 1,3,5-triazine, indole.

Examples of 5- or 6-membered, wholly or partially saturated monocyclic heterocycles include inter alia:

imidazolidine, pyrazolidine, pyrrolidine, piperidine, piperazine, morpholine, thiomorpholine, tetrahydrofuran, tetrahydrothiophene, 1,4-dioxine, imidazoline, pyrazoline, pyrroline, etc.

A. of the compounds of formula I wherein R¹ is (C₄₋₆)cycloalkyl or (C₄₋₆) cycloalkyl(C₁₋₅)alkyl, the following may be mentioned as of particular interest:

Compounds wherein

R³ and R⁴ independently of each other denote (a) hydrogen, (b) C₁₋₈ -alkyl, C₂₋₃ -alkenyl or -alkynyl (wherein the alkyl may be substituted by hydroxy, (C₁₋₄)alkoxy, di(C₁₋₄)alkylamino, furyl, pyrrolidinyl, morpholinyl, pyridinyl or the group ##STR39## wherein B, R⁵ and q are defined as hereinafter) (d) dimethylamino, (f) phenyl, (g) morpholinyl or (h) pyridyl, whilst R³ and R⁴ cannot simultaneously represent hydrogen, dimethylamino or di(C₁₋₄) alkylaminomethyl;

or R³ and R⁴ together with the nitrogen atom to which they are bound denote a pyrrolidinyl, morpholinyl or piperazinyl group, whilst the piperazinyl ring may optionally be N-substituted by unsubstituted phenyl, mono- or di(C₁₋₄)alkoxyphenyl, pyrimidinyl or phenyl(C₁₋₄)alkyl;

more particularly wherein R³ and/or R⁴ denotes unsubstituted phenyl, fluorophenyl, morpholino or 2- or 3-pyridyl;

wherein R³ and/or R⁴ denotes (C₁₋₄)alkyl, preferably methyl or ethyl; or ##STR40## wherein R³ and/or R⁴ denotes (C₂ or C₃)alkyl, which is substituted by hydroxy, methoxy, dimethylamine, furyl, morpholino, pyrrolidinyl or pyridinyl;

wherein R₃ is hydrogen.

Moreover, of these compounds (I), special mention should be made of those wherein R³ is hydrogen and R⁴ is a substituted alkyl of formula VII ##STR41## wherein q' is 0, 1 or 2; R⁶ and R⁷ independently of each other denote hydrogen or (C₁₋₅)alkyl or together with the carbon atom to which they are bound denote a 5- or 6-membered carbocycle;

B denotes phenyl or thienyl;

R⁵ denotes (C₁₋₄)alkyl, halogen (F, Cl, Br, I), hydroxy, (C₁₋₄)alkoxy, amino, thiomethyl, methanesulphonyloxy or methanesulphonamido, or two adjacent R⁵ substituents together denote --O--CH₂ --O-- or --O--CH₂ --CH₂ --O-- and q' denotes 0, 1, 2 or 3, if B is phenyl, and q' denotes 0, 1 or 2 if B is thienyl;

more especially compounds wherein R⁵ denotes (C₁₋₄)alkyl, hydroxy, (C₁₋₄)alkoxy, methanesulphonyloxy or methanesulphonamido, or two adjacent R⁵ substituents together denote --O--CH₂ --O-- or --O--CH₂ --CH₂ --O--;

wherein R⁵ represents hydroxy, (C₁₋₄)alkoxy, methanesulphonyloxy or methanesulphonamido, or two adjacent R⁵ substituents together represent --O--CH₂ --O-- or --O--CH₂ --CH₂ --O--; particularly

wherein R⁵ represents hydroxy, methoxy, methanesulphonyloxy or methanesulphonamido, or two adjacent R⁵ substituents together represent --O--CH₂ --O--; particularly compounds wherein R⁵ is methoxy;

wherein q is zero;

wherein B is phenyl and q is two, preferably wherein the two R⁵ substituents are in positions 2 and 3.

Mention should also be made of compounds wherein R³ and R⁴ together with the nitrogen atom to which they are bound represent morpholino, pyrrolidinyl or piperazinyl (which is N-substituted by methoxyphenyl, phenethyl or 2-pyrimidinyl).

Of the above-mentioned groups of compounds the preferred ones are those wherein

R² represents hydroxy, (C₁₋₄)alkoxy, methanesulphonyloxy or methanesulphonamido, or two adjacent R² substituents together represent --O--CH₂ --O-- or --O--CH₂ --CH₂ --O--; particularly

wherein R² is methoxy.

B. Of the compounds of formula I wherein R¹ is ##STR42## the compounds which may be mentioned as being of particular interest are those wherein

R³ denotes hydrogen; and R⁴ denotes hydrogen, C₃₋₆ -alkenyl; C₃₋₆ -alkynyl; C₃₋₆ -cycloalkyl; C₃₋₆ -cycloalkenyl; straight-chained or branched C₁₋₆ -alkyl, which may optionally be mono- or polysubstituted with the following substituents of groups a) to c), which may be identical or different:

a) halogen; cyano; hydroxy; mercapto; C₁₋₄ -alkoxy; C₁₋₄ -alkylthio; amino; mono-C₁₋₄ -alkylamino; di-C₁₋₄ -alkylamino (wherein the alkyl radicals may be identical or different), phenoxy (wherein the phenyl group may be substituted as in (b)),

b) phenyl; optionally mono- or polysubstituted (with identical or different substituents) by the groups halogen, trifluoromethyl, C₁₋₄ -alkoxy, hydroxy, mercapto, C₁₋₄ -alkylthio, C₁₋₄ -alkyl, amino, mono-C₁₋₄ -alkylamino, di-C₁₋₄ -alkylamino (wherein the alkyl groups may be identical or different), C₂₋₃ -acylamino, C₂₋₃ -acyloxy and the --O--CH₂ --O-- or --O--(CH₂)₂ --O-- group vicinally bound to the phenyl system,

C) a 5- or 6-membered saturated or wholly or partially unsaturated monocyclic heterocycle having up to 3 heteroatoms selected from the group comprising N, O and S; and as a bicyclic heterocycle indole (whilst the above-mentioned heterocycles may be mono- or polysubstituted by C₁₋₄ -alkyl), C₃₋₆ -cycloalkyl; C₅ - or C₆ -cycloalkenyl; C₂₋₃ -acyl; C₁₋₄ -alkylsulphonyl; or phenyl (which may in turn be substituted up to three times as described under b);

or R³ is hydrogen and R⁴ is phenyl which may be substituted as specified under b) above;

R₃ and R₄ independently of each other denote C₁₋₄ -alkyl, which may optionally be phenyl-substituted, whilst the phenyl substituent may in turn be substituted as under b) hereinbefore;

or

R³ and R⁴ together with the nitrogen atom to which they are bound denote a wholly or partially saturated heterocyclic 5- or 6-membered ring (which may also contain up to two further heteroatoms selected from the group N, O, S), whilst the heterocyclic group thus obtained may be substituted by C₁₋₄ -alkyl, hydroxy, phenyl or benzyl (whilst this phenyl group or the phenyl group of the benzyl group is substituted as under b) hereinbefore).

Within the description of general formula I is included in particular 3,4-dihydroisoquinoline derivatives wherein NR³ R⁴ is ##STR43## wherein X¹ represents phenyl mono- or disubstituted by trifluoromethyl or ethoxy, phenyl substituted by methoxy and fluorine or 2-methoxyphenyl,

X² is --CH₂ --CH₂ -- or

--CH₂ --CH(CH₃)--,

and

X³ is 2,3,4-trimethoxyphenyl, 2,3-dimethoxyphenyl, 2,6-dimethoxyphenyl, 3,6-dimethoxyphenyl, thienyl, phenyl mono- or disubstituted by trifluoromethyl or ethoxy, or phenyl substituted by methoxy and fluorine,

or the pharmaceutically acceptable salts thereof.

The preferred compounds (I) are those wherein NR³ R⁴ is ##STR44##

Carbocyclically and heterocyclically anellated dihydropyridines of the formula ##STR45## and the tautomeric forms thereof, wherein R³ denotes hydrogen and

R⁴ denotes hydrogen; straight-chained or branched unsubstituted C₁₋₅ -alkyl; allyl; propargyl; C₃₋₆ -cycloalkyl; 3-chlorophenyl; 2-methyl-3-chlorophenyl; or C₁₋₃ -alkyl, which is monosubstituted with one of the substituents of groups d) to f) listed below;

d) cyano, hydroxy, methoxy, dimethylamino

e) phenyl, 3,4-methylenedioxyphenyl, phenyl substituted by one, two or 3 methoxy groups, 3-hydroxy-4-methoxyphenyl,

f) morpholino, pyridin-2-yl, indol-3-yl, furan-2-yl, thiophen-2-yl, pyridin-3-yl, pyridin-4-yl;

R³ and R⁴ independently of each other denote methyl; ethyl; 3-cyanopropyl; benzyl; or 3,4,5-trimethoxyphenethyl or

R³ and R⁴ together with the nitrogen atom to which they are bound represent morpholine; thiomorpholine; pyrrolidine; piperazine; 4-methylpiperazine; 4-benzylpiperazine or 4-(2-methoxyphenyl)piperazine;

and

A represents the anellated ring systems ##STR46## are preferred.

The phenyl group of the compound of general formula I may contain 1, 2 or 3 R substituents. These R substituents may be identical or different from one another. Preferably R is C₁₄ -alkyl (preferably methyl), halogen (preferably fluorine, chlorine or bromine) or CF₃.

C. Of the compounds of formula I wherein R¹ is unsubstituted phenyl, particular mention may be made of those compounds wherein

R³ is hydrogen and

R⁴ is hydrogen, C₃₋₆ -alkenyl; C₃₋₆ -alkynyl-; C₃₋₆ -cycloalkyl; C₃₋₆ -alkyl anellated with a benzo group; C₃₋₆ -cycloalkenyl; straight-chained or branched C₁₋₁₀ -alkyl, which may optionally be mono- or polysubstituted with the following substituents of groups a) to c), which may be identical or different:

a) cyano; hydroxy; C₁₋₄ -alkoxy; C₁₋₄ -alkylthio; C₁₋₄ -alkyloxycarbonyl; amino; mono-C₁₋₄ -alkylamino; di-C₁₋₄ -alkylamino (wherein the alkyl radicals may be identical or different), phenoxy (wherein the phenyl group may be substituted as under (b); naphthoxy;

b) phenyl; optionally mono-, di- or trisubstituted (with identical or different substituents) by the groups halogen, trifluoromethyl, C₁₋₄ -alkoxy, hydroxy, mercapto, C₁₋₄ -alkylthio, C₁₋₄ -alkyl, azido, amino, mono-C₁₋₄ -alkylamino, di-C₁₋₄ -alkylamino (wherein the alkyl groups may be identical or different), C₂₋₃ -acylamino, C₂₋₃ -acyloxy and the group --O--CH₂ --O-- or --O--(CH₂)₂ --O-- vicinally bound to the phenyl system;

C) a 5- or 6-membered saturated or wholly or partially unsaturated monocyclic heterocycle having up to 3 heteroatoms from the group comprising N, O and S; and as a bicyclic heterocycle indole (whilst the above-mentioned heterocycles may be mono- or polysubstituted by C₁₋₄ -alkyl)

C₃₋₁₀ -cycloalkyl (optionally bridged cycloalkyl); C₅ - or C₆ -cycloalkenyl; C₂₋₃ -acyl; C₁₋₄ -alkylsulphonyl;

or R⁴ is phenyl which may be substituted as specified under (b) hereinbefore;

R³ and R⁴ independently of each other are each defined as for R⁴ above, and preferably represent C₁₋₆ -alkyl, which may optionally be phenyl-substituted, whilst the phenyl substituent may in turn be substituted as under b) hereinbefore;

or

R³ and R⁴ together with the nitrogen atom to which they are bound represent a wholly or partially saturated heterocyclic 5- or 6-membered ring (which may also contain up to 2 further heteroatoms selected from the group comprising N, O and S), whilst the heterocyclic group thus obtained may be substituted by C₁₋₄ -alkyl, hydroxy or (CH₂)_(q") R⁸ (where q"=0,1,2,3 or 4);

and

R⁸ denotes a phenyl radical or phenoxy radical wherein, if desired, the phenyl group is substituted as under b) hereinbefore, or R⁸ denotes naphthoxy;

A denotes the anellated ring systems ##STR47## wherein R² is as hereinbefore defined, more particularly wherein R³ is hydrogen and

R⁴ is hydrogen; C₃₋₆ -alkenyl; C₃₋₆ -alkynyl; C₃₋₆ -cycloalkyl; C₃₋₆ -cycloalkenyl; straight-chained or branched C₁₋₆ -alkyl which may optionally be mono- or polysubstituted with the following substituents of groups a) to c), which may be identical or different:

a) halogen; cyano; hydroxy; mercapto; C₁₋₄ -alkoxy; C₁₋₄ -alkylthio; amino; mono-C₁₋₄ -alkylamino; di-C₁₋₄ -alkylamino (wherein the alkyl radicals may be identical or different), phenoxy (wherein the phenyl group may be substituted as under (b),

b) phenyl; optionally mono- or polysubstituted (with identical or different substituents) by the groups halogen, trifluoromethyl, C₁₋₄ -alkoxy, hydroxy, mercapto, C₁₋₄ -alkylthio, C₁₋₄ -alkyl, amino, mono-C₁₋₄ -alkylamino, di-C₁₋₄ -alkylamino (wherein the alkyl groups may be identical or different), C₂₋₃ -acylamino, C₂₋₃ -acyloxy and the group --O--CH₂ --O-- or --O--(CH₂) ₂ --O-- vicinally bound to the phenyl system

C) a 5- or 6-membered saturated or wholly or partially unsaturated monocyclic heterocycle having up to 3 heteroatoms from the group N, O, S; and as a bicyclic heterocycle indole (whilst the above-mentioned heterocycles may be mono- or polysubstituted by C₁₋₄ -alkyl), C₃₋₆ -cycloalkyl; C₅ - or C₆ -cycloalkenyl; C₂₋₃ -acyl; C₁₋₄ -alkylsulphonyl; or phenyl (which may in turn be substituted up to three times as described under b);

R³ is hydrogen and

R⁴ is phenyl which may be substituted as specified under (b) hereinbefore;

R³ and R⁴ independently of each other denote C₁₋₄ -alkyl, which may optionally be phenyl-substituted, whilst the phenyl substituent may in turn be substituted as under b) hereinbefore;

or

R³ and R⁴ together with the nitrogen atom to which they are bound denote a wholly or partially saturated, heterocyclic 5- or 6-membered ring (which may also contain up to 2 further heteroatoms from the group N, O, S), whilst the resulting heterocycle may be substituted by C₁₋₄ -alkyl, hydroxy or (CH₂)_(q") -R⁸ (wherein q"=0 or 1)

and

R⁸ denotes a phenyl radical which may optionally be substituted as under b) hereinbefore;

A denotes the anellated ring systems ##STR48## wherein R² is as hereinbefore defined.

General formula I as described covers, in particular, 3,4-dihydroisoquinoline derivatives of the general formula ##STR49## wherein NR³ R⁴ represents ##STR50## wherein X¹ denotes phenyl which is mono- or disubstituted by trifluoromethyl or ethoxy, phenyl which is substituted by methoxy and fluorine, or 2-methoxyphenyl,

X² denotes --CH₂ --CH₂ -- or

--CH₂ --CH(CH₃)--,

and

X³ denotes 2,3,4-trimethoxyphenyl, 2,3-dimethoxyphenyl, 2,6-dimethoxyphenyl, 3,6-dimethoxyphenyl, thienyl, phenyl which is mono- or disubstituted by trifluoromethyl or ethoxy, or phenyl substituted by methoxy and fluorine,

or the pharmaceutically acceptable salts thereof.

Particularly preferred compounds are those wherein NR³ R⁴ denotes ##STR51## wherein X¹ is 2-methoxyphenyl, which may additionally be substituted by fluoro,

X² is --CH₂ --CH₂ -- and

X³ is 2,3,4-trimethoxyphenyl, 2,3-dimethoxyphenyl, 2,6-dimethoxyphenyl, 3,6-dimethoxyphenyl, 2- or 3-thienyl, phenyl substituted by trifluoromethyl or ethoxy, or phenyl substituted by methoxy and fluorine.

Particular mention should be made of compounds of the formula ##STR52## and the tautomeric forms thereof wherein R³ is hydrogen and

R⁴ is hydrogen; straight-chained or branched unsubstituted C₁₋₅ -alkyl; allyl; propargyl; C₃₋₆ -cycloalkyl; 3-chlorophenyl; 2-methyl-3-chlorophenyl; or C₁₋₃ -alkyl, which is monosubstituted with one of the substituents of groups d) to f) listed hereinafter;

d) cyano, hydroxy, methoxy, dimethylamino

e) phenyl, 3,4-methylenedioxyphenyl, phenyl substituted by one, two or 3 methoxy groups, 3-hydroxy-4-methoxyphenyl,

f) morpholino, pyridin-2-yl, indol-3-yl, furan-2-yl, thiophen-2-yl, pyridin-3-yl, pyridin-4-yl

R³ and R⁴ independently of each other denote methyl; ethyl; 3-cyanopropyl; benzyl; or 3,4,5-trimethoxyphenethyl or

R³ and R⁴ together with the nitrogen atom to which they are bound denote morpholine; thiomorpholine; pyrrolidine; piperazine; 4-methylpiperazine; 4-benzylpiperazine; or 4-(2-methoxyphenyl)piperazine;

and ##STR53## denotes the anellated ring systems ##STR54## wherein R² is as hereinbefore defined.

The compounds of formula I may be prepared by methods known per se, preferably according to the method described in German Patent Application P 37 18 570.5, EP 358 957, EP 37 934 and EP 251 794.

In the presence of a condensing agent a malonic acid amide of general formula IV ##STR55## wherein R¹, R², R³, R⁴ and m are as hereinbefore defined and Ar denotes phenyl or 2- or 3-thienyl, may be cyclised into the corresponding compounds.

Suitable condensing agents for this process include strong Lewis acids such as phosphorusoxychloride, phosphoruspentachloride, phosphorustrichloride, phosphoruspentoxide, titanium tetrachloride, boron trifluoride, tin tetrachloride, and also inorganic acids such as polyphosphoric acid, sulphuric acid, fluorosulphonic acid and hydrofluoric acid, or mixtures of condensing agents such as a mixture of phosphorusoxychloride and phosphoruspentachloride, or a mixture of phosphoruspentoxide and (C₁₋₄)alkylsulphonic acid, e.g. with a P₂ O₅ -content of about 10% by weight.

The cyclisation may be carried out in the presence or absence of a solvent. All inert solvents are suitable provided that they have sufficient solubility for the reactants and a high enough boiling point, e.g. benzene, alkylbenzenes (e.g. toluene, xylene), chlorobenzenes, chloroform, acetonitrile and decalin. According to a preferred alternative embodiment of the process the condensing agent, e.g. phosphorusoxychloride or a (C₁₋₄)alkylsulphonic acid/phosphoruspentoxide mixture is used without the addition of solvents.

Preferably, the cyclisation is carried out using phosphorusoxychloride or, in difficult cases, with a mixture of phosphoruspentoxide and (C₁₋₄)alkylsulphonic acid (preferably methanesulphonic acid). The reaction may be carried out within a wide temperature range, preferably with warming or heating to 50° C. up to about the boiling point of the reaction mixture.

The reaction time required will range from 2 to 15 hours depending on the starting compound of formula IV.

The compounds of formula I are bases and may be converted in the usual way with organic or inorganic acids and salt-forming agents and complexing agents into any desired physiologically acceptable adducts (salts).

Examples of acids suitable for salt formation include hydrochloric acid, hydrobromic acid, hydriodic acid, hydrofluoric acid, sulphuric acid, phosphoric acid, nitric acid, acetic acid, propionic acid, butyric acid, caproic acid, valeric acid, oxalic acid, malonic acid, succinic acid, maleic acid, fumaric acid, lactic acid, tartaric acid, citric acid, malic acid, benzoic acid, p-hydroxybenzoic acid, phthalic acid, cinnamic acid, salicylic acid, ascorbic acid, methanesulphonic acid and the like.

The compounds may be given orally, parenterally or topically. The desired therapeutic dose depends on the indication and form of preparation and can be determined experimentally. Suitable forms include, for example, tablets, capsules, suppositories, solutions, syrups, emulsions, aerosols or dispersible powders. Tablets may be produced, for example, by mixing the active substance or substances with known excipients, e.g. inert diluents such as calcium carbonate, calcium phosphate or lactose, disintegrants such as corn starch or alginic acid, binders such as starch or gelatine, lubricants such as magnesium stearate or talc and/or agents for obtaining delayed release, such as carboxypolymethylene, carboxymethylcellulose, cellulose acetate phthalate or polyvinylacetate. The tablets may also consist of several layers.

Coated tablets may be produced analogously by coating cores made in the same way as the tablets with substances conventionally used for tablet coatings, e.g. collidone or shellack, gum arabic, talc, titanium dioxide or sugar. In order to obtain delayed release or avoid incompatibilities, the core may also consist of several layers. Similarly, the tablet coating may consist of several layers to achieve delayed release, whilst the excipients mentioned for the tablets may be used.

Syrups containing the active substances or combinations of active substances according to the invention additionally contain a sweetener such as saccharin, cyclamate, glycerol or sugar as well as a flavour enhancer, e.g. a flavouring such as vanillin or orange extract. They may also contain suspension adjuvants or thickeners such as sodium carboxymethylcellulose, wetting agents, e.g. condensation products of fatty alcohols with ethylene oxide or preservatives such as p-hydroxybenzoates.

Injectable solutions are produced in the usual way, e.g. by adding preservatives such as p-hydroxybenzoates or stabilisers such as alkali metal salts of ethylene diamine tetraacetic acid, and are then transferred into injection vials or ampoules.

Capsules containing one or more active substances or combinations of active substances may be prepared for example by mixing the active substances with inert carriers such as lactose or sorbitol and encapsulating them in gelatine capsules.

Suitable suppositories may be produced for example by mixing with carriers provided for this purpose, such as neutral fats or polyethyleneglycol or derivatives thereof.

The compounds may be given both enterally and parenterally. The proposed dose for oral administration is 0.1 to 500 mg of active substance per dose, and for intravenous use 0.05 to 150 mg per dose. The desired therapeutic dose depends on the indication and form of preparation and can be determined experimentally.

The pharmaceutical compositions are suitable for oral or parenteral and possibly topical use. The pharmaceutical forms are predominantly plain or coated tablets, ampoules and syrups. The individual dose of these preparations is between 1.0 and 200 mg, preferably 20 to 50 mg per 75 kg of body weight. Depending on the severity of the case, 1 to 3 single doses are generally administered per day.

The Examples which follow are intended to illustrate the invention:

EXAMPLE 1 3,4-Dihydro-1-benzyl-6,7-dimethoxy-α- di-2-(2,3,4-trimethoxyphenyl)ethyl!aminocarbonyl-isoquinoline-hydrochloride ##STR56##

a) 2-(3,4-Dimethoxyphenyl)ethylaminocarbonyl-phenyl-acetic acid-N,N-di- 2-(2,3,4-trimethoxyphenyl)-ethyl!amide

To a solution of 18.0 g (52.4 mmol) of monoethylphenylmalonate-2-(3,4-dimethoxyphenyl)ethylamide in 150 ml of anhydrous dimethylformamide are added, at ambient temperature 9.0 g (55.5 mmol) of N,N'-carbonyldiimidazole in batches. After 30 minutes 18.0 g (44.3 mmol) of di- 2-(2,3,4-trimethoxyphenyl)ethyl!amine are added and the mixture is stirred for 30 minutes. Then the solvent is distilled off in vacuo, the residue is taken up in 1.5 liters of CH₂ Cl₂ and extracted twice with 250 ml of water and 200 ml of 1N HCl, one after the other. The organic phase is evaporated down after drying over Na₂ SO₄ and, after purification over a silica gel column (eluant: CH₂ Cl₂ /MeOH 100:2) the residue is crystallised from ethyl acetate/ether.

Yield: 35.5 g

b) 35.0 g (47.5 mmol) of amide (from step a) and 15 ml (164 mmol) of phosphorusoxychloride are heated to boiling for 30 minutes in 150 ml of anhydrous acetonitrile. After the reaction has ended (monitored by thin layer chromatography) the solvent and any unused phosphorusoxychloride are distilled off in vacuo. The residue is mixed with ice water, made alkaline with soda solution and extracted in batches with about 1 liter of CH₂ Cl₂. The organic phase is washed with water, dried over Na₂ SO₄ and evaporated down. The residue is purified twice over a silica gel column (1st eluant: CH₂ Cl₂ :MeOH 100:2→100:4 rising; 2nd eluant: CH₂ Cl₂ /ethyl acetate 1:1).

The hydrochloride is formed from the purified product (6.5 g) by dissolving in about 50 ml of ethanol and adding alcoholic hydrochloric acid. After evaporation and drying under a high vacuum at 50° C. 11.5 g of the desired product remain.

(M.p. 56-64° C., amorphous)

EXAMPLE 2 2-Phenylmalonic acid-N-(2-(3,4-dimethoxyphenyl)ethyl)-N',N'-di-(2-(2-fluorophenyl)ethyl)-diamide

To a solution of 17.2 g (0.05 mol) of phenylmalonic acid N-(2-(3,4-dimethoxyphenyl)ethyl)-monoamide in 200 ml of anhydrous CH₂ Cl₂, 8.1 g (0.05 mol) of N,N'-carbonyldiimidazole are added in batches at ambient temperature and with stirring. After about 30 minutes a suspension consisting of 14.9 g (0.05 mol) of di-(2-(2-fluorophenyl)ethyl)amine-hydrochloride and 5.1 g=6.4 ml (0.05 mol) of triethylamine in 100 ml of anhydrous CH₂ Cl₂ is stirred into the reaction mixture. After 15 hours' stirring, 200 ml of water are added, the mixture is acidified with dilute hydrochloric acid, the organic phase is separated off and the aqueous phase is extracted 3 times with 100 ml of CH₂ Cl₂. The combined organic phases are dried over Na₂ SO₄ and evaporated down. R,S (3,4-Dihydro-6,7-dimethoxyisoquinolin-1-yl)-2-phenyl N,N-di-(2-(2-fluorophenyl)ethyl-acetamide-oxalate

A mixture of 29.8 g (0.049 mol) of amide, 150 ml of anhydrous CH₂ C₂ and 17.5 g=9.4 ml (0.10 mol) of POCl₃ is refluxed for 10 hours. After the reaction has ended (TLC monitoring) the mixture is stirred into a combination of 400 ml of ice water and 200 ml of CH₂ Cl₂. Then it is neutralised with a saturated soda solution, the organic phase is separated off, the aqueous phase is extracted 3 times with 100 ml of CH₂ Cl₂. The combined phases are dried over Na₂ SO₄ and evaporated down in vacuo. The residue is purified over silicic acid (eluant: CH₂ Cl₂ =methanol=100:2) and then dissolved in a little ethanol and converted into the oxalate by adding a stoichiometric quantity of an alcoholic oxalic acid solution. The oxalate is also crystallised by the addition of ether. M.p.: 144-146° C.

EXAMPLE 3 (R,S)-(3,4-Dihydro-6,7-dimethoxyisoquinolin-1-yl)-2-(4-methoxyphenyl)-N N-di(2-(2,3,4-trimethoxyphenyl)ethyl)-acetamide-oxalate

Ingredients:

5 g (6.57 mMol) of 4-methoxyphenylmalonic acid-N-(2-(3,4-dimethoxyphenyl)ethyl)-N'-(2-(2,3,4-trimethoxyphenyl)-ethyl-diamide ("diamide") 18 ml of acetonitrile, 3.02 g (19.7 mMol) of phosphorusoxychloride, 600 mg (6.66 mmol) of oxalic acid, anhydrous, 200 ml of ether

Method: The reaction mixture of the "diamide", acetonitrile and phosphorusoxychloride is refluxed for 1 hour under protective N₂. After cooling with ice water it is diluted with 100 ml of ethyl acetate and then washed twice each with ice water, 50 ml of saturated NaHCO₃ solution, water and saturated NaCl solution. The organic phase is dried over MgSO₄ and poured, with stirring, into a solution of 706 mg (7.84 mMol) of anhydrous oxalic acid in 200 ml of absolute ether.

The reaction product was initially precipitated as an oil as the oxalic acid salt, which crystallised after standing for some time with stirring. After standing overnight in the refrigerator it was suction filtered, washed with ether and dried. M.p.: 107-110° C. The structure was confirmed by NMR spectroscopy.

R_(f) values: 0.53 (ethyl acetate)

R_(f) values: 0.6 (acetonitrile:H₂ O=9:1)

Preparation of the starting compound:

Part A

Diethyl 4-methoxyphenylmalonate

Ingredients:

150 ml of absolute ethanol

7.5 g (0.33 mol) of sodium

300 ml of diethylcarbonate

62.5 g (0.3 mol) of ethyl 4-methoxyphenylacetate

Method:

Sodium is dissolved in absolute ethanol and evaporated to dryness in vacuo. Whilst cooling with ice water the residue is combined with diethylcarbonate and ethyl 3-chlorophenylacetate, with stirring. Then the ethanol is slowly (2-3 hours) distilled off through a 40 cm column (Raschig rings) under a high vacuum at 40-70° C. and 200 mM. After cooling, the mixture is acidified with 30 ml of glacial acetic acid and mixed with 150 ml of water. The oil precipitated is washed successively with water and saturated NaCl solution and dried over MgSO₄. The residue is distilled in a bulb tube using an oil pump; b.p.: 0.5 mM: 150-155° C.

Monoethyl 4-methoxyphenylmalonate

Ingredients:

52.2 g (0.196 mol) of diethylester

120 ml of ethanol

120 ml of water

12.3 g (0.22 mol) of KOH in 60 ml of water and 60 ml of ethanol

Method:

An ethanolic solution of diethyl 4-methoxyphenylmalonate is mixed with the aqueous alcoholic KOH solution, with stirring and cooling with ice, and stirred for 75 minutes. Then it is acidified with a saturated citric acid solution and extracted 3 times with methylene chloride. The organic phase is washed successively with water and saturated NaCl solution, dried over MgSO₄ and the solvent is distilled off in vacuo. The crystalline residue is recrystallised from methylene chloride/petroleum ether (40-800).

Yield: 34.4 g (73.6% of theory).

The structure was confirmed by NMR spectroscopy.

Monoethyl N-(2,3,4-dimethoxyphenyl)ethyl)-4-methoxy-phenyl-malonate-amide

Ingredients:

34.4 g (0.144 mol) of monoethyl 4-methoxy-phenylmalonate

150 ml of anhydrous tetrahydrofuran

23.3 g (0.144 mol) of N,N'-carbonyldiimidazole

26.1 g (0.144 mol) of 2-(3,4-dimethoxyphenyl)ethylamine in 50 ml of anhydrous tetrahydrofuran

Method:

The carbonyldiimidazole is added in batches at 50° C., with stirring, to the solution of the 4-methoxy-phenylmalonic acid hemiester in THF. After 30 minutes' stirring at ambient temperature the amine is added whilst cooling with ice and the mixture is stirred for 16 hours at ambient temperature. The reaction mixture is evaporated down and the residue is taken up in CH₂ Cl₂. It is washed twice with water, then 10% KHSO₄ solution, saturated NaHCO₃ solution, water and saturated saline solution. After drying over MgSO₄ the organic phase is evaporated down and the oily residue (7.7 g) is crystallised from ethyl acetate.

4-Methoxy-phenylmalonic acid-N-(2(3,4-dimethoxyphenyl)-ethyl)-amide

Ingredients:

44.16 g (0.11 mol) (4-methoxy-phenylmalonic acid hemiester amide)

300 ml methanol

120 ml (0.12 mol) 1N sodium hydroxide solution

Method:

At 5-10° C. the sodium hydroxide solution is stirred into the methanolic solution of the hemiester amide for 30 minutes. After 3 hours' stirring at ambient temperature it is evaporated down, diluted with water, then extracted twice with CH₃ Cl. The aqueous phase is adjusted to pH 1 with concentrated HCl and extracted twice with CH₃ Cl. After washing with saturated saline solution the organic phase is dried over MgSO₄ and evaporated down. The crystalline residue is crystallised from CH₃ cl.

Part B

2-(2,3,4-Trimethoxyphenyl)-1-nitro-ethers

    ______________________________________     Ingredients:     ______________________________________     400     g     (2.04 mol)                            of 2,3,4-trimethoxybenzaldehyde     1740    g              of glacial acetic acid     172.6   g     (2.24 mol)                            of anhydrous ammonium acetate     656     ml             of nitromethane     ______________________________________

Method:

The mixture of benzaldehyde, ammonium acetate, nitromethane and glacial acetic acid is stirred for 30 minutes at boiling temperature under nitrogen protection. The mixture is cooled to -5° C. and poured into 5 liters of ice water with stirring. The viscous residue is extracted exhaustively with CH₂ Cl₂. The organic phase is washed with water, dried over MgSO₄ and evaporated down. The brown oily residue (510 g) crystallises on standing. It is dissolved in 470 ml of ethyl acetate and crystallised by the addition of 3.9 liters of cyclohexane.

2-(2,3,4-Trimethoxyphenyl)ethylamine-hydrochloride

    ______________________________________     Ingredients:     ______________________________________     1.445    g     (0.60 mol)                             "Nitrostyrol"     1.9      l              of water     232      ml             of conc. hydrochloric acid                             (analytical grade)     88       g              PdC (10%)     ______________________________________

Method:

The above reaction mixture is hydrogenated at 60° C. under 35 bar for 2.25 hours. Then it is evaporated down in vacuo, the residue is taken up in ethanol and evaporated to dryness. It is then dissolved in ethanol and the reaction product is crystallised by the addition of ether.

2,3,4-Trimethoxybenzylalcohol

    ______________________________________     Ingredients:     ______________________________________     500      g     (2.55 mol)                             2,3,4-trimethoxybenzaldehyde     5.0      l              of methanol     13.0     g              of PtO.sub.2     ______________________________________

Method:

The reduction is carried out at 20° C. under 5 bar and is complete after 30 minutes. After the solvent has been evaporated off the residue (501.5 g) is distilled in a high vacuum (B.p. 116°; 0.5 mbar).

2,3,4-Trimethoxybenzylchloride

    ______________________________________     Ingredients:     ______________________________________     50.0    g     (0.25 mol)                            2,3,4-trimethoxybenzylalcohol     800     ml             of anhydrous methylene chloride     59.4    g     (0.5 mol)                            of thionylchloride     ______________________________________

Method:

The solution of the alcohol in anhydrous CH₂ Cl₂ is slowly mixed with SOCl₂ with stirring and cooling with ice and common salt. The mixture is stirred for a further 15 minutes in the cold, then for 2 hours at ambient temperature. The solvent and excess thionylchloride are eliminated in vacuo, the residue is taken up in CH₂ Cl₂ and shaken successively with saturated NaHCO₃ solution, water and saturated saline solution. After drying over MgSO₄ the solvent is removed in vacuo and the residue is distilled in a bulb tube furnace using an oil pump (b.p. 118° C.; 0.1 mbar).

2,3,4-Trimethoxybenzylcyanide

    ______________________________________     Ingredients:     ______________________________________     75.8    g     (0.35 mol)                             2,3,4-trimethoxybenzylchloride     700     ml              of anhydrous acetone     3.45    g     (0.023 mol)                             of NaI     25.7    g     (0.53 mol)                             of dried powdered NaCN     ______________________________________

Method:

The reaction mixture consisting of benzyl chloride, NaI and NaCN in anhydrous acetone is stirred for 20 hours at boiling temperature. After cooling it is suction filtered and the solvent is eliminated in vacuo. The residue is dissolved in ethyl acetate, shaken first with water then with saturated saline solution and dried over MgSO₄. After removal of the solvent the residue is distilled in a bulb tube furnace at 0.015 mbar (b.p. 135°C.).

2,3,4-Trimethoxyphenylacetic acid

    ______________________________________     Ingredients:     ______________________________________     138.5  g     (0.67 mol)                           2,3,4-trimethoxybenzylcyanide,     53.5   g     (1.34 mol)                           NaOH dissolved in 215 ml of water     ______________________________________

Method:

The mixture of benzyl cyanide and aqueous sodium hydroxide solution is refluxed for 7 hours, then after cooling acidified with 6N H₂ SO₄ and extracted three times with CH₂ Cl₂. The organic phase is washed with water and saturated NaCl solution and dried over MgSO₄. After removal of the solvent the residue is dissolved in 200 ml of CH₂ Cl₂ and crystallised by the addition of 1500 ml of cyclohexane.

2,3,4-Trimethoxyphenyl-N-(2-(2,3,4-trimethoxy-phenyl)ethyl)-acetamide

    ______________________________________     Ingredients:     ______________________________________     72.4    g     (0.32 mol)                            2,3,4-trimethoxyphenylacetic acid     400     ml             anhydrous tetrahydrofuran     51.8    g     (0.32 mol)                            N,N'-carbonyldiimidazole (CDI)     79.3    g     (0.32 mol)                            2-(2,3,4-trimethoxyphenyl)-                            ethylamine-hydrochloride     500     ml             anhydrous tetrahydrofuran     32.4    ml    (0.32 mol)                            triethylamine     ______________________________________

Method:

The phenylacetic acid dissolved in anhydrous THF is converted into the imidazolide with stirring at 5° C. by the batch-wise addition of CDI. After 30 minutes a suspension of amine hydrochloride, triethylamine in anhydrous THF is stirred in at ambient temperature. After 16 hours the mixture is evaporated down in vacuo and the residue is divided between CH₂ Cl₂ and 2N HCl. Then it is washed successively with water, saturated NaHCO₃ solution, water and saturated NaCl solution. After drying over MgSO₄ the organic phase is evaporated down, the residue is dissolved in 200 ml of ethyl acetate and the product is crystallised by the addition of 700 ml of cyclohexane.

Di-2-(2,3,4-trimethoxyphenyl)ethylamine-hydrochloride

    ______________________________________     Ingredients:     ______________________________________     113.4   g     (0.27 mol)                            "phenylacetic acid amide"     470     ml             anhydrous tetrahydrofuran     270     ml    (0.54 mol)                            BH.sub.3.S(CH.sub.3).sub.2  in THF (2     ______________________________________                            mol/l)

Method:

At 65° C. the borane complex mixture is added dropwise to the solution of the acid amide in anhydrous THF under protective N₂. After the addition has ended the mixture is stirred for a further 15 minutes at 65° C. Then the reaction mixture is cooled to 5° C. It is carefully acidified with methanolic hydrochloric acid, evaporated down in vacuo (drawn off) and the residue is crystallised from ethanol with the addition of ether.

Part C

4-Methoxy-phenylmalonic acid-N-(2-(3,4-dimethoxyphenyl)-ethyl)-N'-(2-(2,3,4-trimethoxyphenyl)ethyl)-di-amide

    ______________________________________     Ingredients:     ______________________________________     3.73   g     (10 mMol)                           4-methoxy-phenylmalonic acid hemiamide     25     ml             anhydrous THF     1.62   g     (10 mMol)                           N,N'-carbonyldiimidazole     4.1    g     (10 mMol)                           di-2-(2,3,4-trimethoxyphenyl)ethylamine-                           hydrochloride     40     ml             anhydrous THF     1.01   g     (10 mMol)                           triethylamine = 1.39 ml     ______________________________________

Method:

The carbonyldiimidazole is added in batches to the solution of the monoamide in THF at 5° C. with stirring. The reaction mixture is stirred for 30 minutes at ambient temperature. Then, whilst cooling with ice, it is reacted with a suspension of the amine-hydrochloride in THF and triethylamine. After 16 hours' stirring at ambient temperature it is evaporated down and the residue is dissolved in ethyl acetate. The organic phase is washed successively with water, 5% KHSO₄ solution, saturated NaHCO₃ solution, water and unsaturated NaCl solution. After drying over MgSO₄ the mixture is evaporated down, the residue (7.2 g) is purified on 210 g of silica gel (eluant: ethyl acetate/n-hexane=2:1).

The following Tables contain examples of compounds according to the invention, whilst Tables 10 to 20 list new compounds.

                  TABLE 1     ______________________________________     Structural type:      ##STR57##     No.  X                     Structure                                         Salt form     ______________________________________     1.   OCH.sub.3             II       --     2.   OC.sub.2 H.sub.5      II       --     3.   NHCH.sub.3            I        Cl     4.   NHC.sub.2 H.sub.5     I        --     5.   NH(CH.sub.2).sub.2CH.sub.3                                I        Cl     6.   NH(CH.sub.2).sub.3CH.sub.3                                I        Cl     7.   NH(CH.sub.2).sub.4CH.sub.3                                I        Cl     8.   NHCH(CH.sub.3).sub.2  I        Cl     9.   NHCH.sub.2CH(CH.sub.3).sub.2                                I        Cl     10.  NH(CH.sub.2).sub.2CH(CH.sub.3).sub.2                                I        Cl     11.  NHC(CH.sub.3).sub.3   I        Cl     12.  NHCH(CH.sub.3)C.sub.2 H.sub.5                                I        Cl     13.  NHCH.sub.2CHCH.sub.2  I        Cl     14.  NHCH.sub.2CCH         I        Cl     15.  NH(CH.sub.2).sub.2OH  II       Cl     16.  NHCH.sub.2CH(OH)CH.sub.3                                I        Cl     17.  NH(CH.sub.2).sub.2OCH.sub.3                                II       Cl     18.  NH(CH.sub.2).sub.3OCH.sub.3                                II       Cl     19.  NH(CH.sub.2).sub.2N(CH.sub.3).sub.2                                I        Cl.sub.2     20.  NH(CH.sub.2).sub.3N(CH.sub.3).sub.2                                II       Cl.sub.2           ##STR58##            II       --           ##STR59##            I        Cl           ##STR60##            I        --           ##STR61##            I        --           ##STR62##            I        --           ##STR63##            II       --           ##STR64##            II       Cl           ##STR65##            I        --           ##STR66##            I        --     30.           ##STR67##            II       --           ##STR68##            I        --           ##STR69##            II       --           ##STR70##            I        Cl           ##STR71##            I        Cl           ##STR72##            II       --     36.  N(CH.sub.3).sub.2     I        Cl     37.  N(C.sub.2 H.sub.5).sub.2                                I        Cl           ##STR73##            I        Cl     38a.           ##STR74##            I        Cl           ##STR75##            I        Cl     40.           ##STR76##            I        --           ##STR77##            I II     --           ##STR78##            II       Cl.sub.2           ##STR79##            I        --           ##STR80##            I        Cl     ______________________________________

                  TABLE 2     ______________________________________     Structural type:      ##STR81##     No.  X                     Structure                                         Salt form     ______________________________________     45.  OC.sub.2 H.sub.5      II       --           ##STR82##            II       Cl           ##STR83##            II       --           ##STR84##            II       --           ##STR85##            II       --     50.           ##STR86##            I        --     ______________________________________

                  TABLE 3     ______________________________________     Structural type:      ##STR87##     No.   X                    Structure                                         Salt form     ______________________________________     51.   OC.sub.2 H.sub.5     I        --                                II     52.   NH(CH.sub.2).sub.3CH.sub.3                                I        --     53.   NH(CH.sub.2).sub.4CH.sub.3                                I        --     54.   NHCH(CH.sub.3).sub.2 I        Cl     55.   NHCH.sub.2CH(CH.sub.3).sub.2                                I        --     56.   NHCH.sub.2CHCH.sub.2 I        --            ##STR88##           I        Cl            ##STR89##           I        Cl            ##STR90##           I     60.            ##STR91##           I            ##STR92##           I        --            ##STR93##           I        --            ##STR94##           I        Cl            ##STR95##           I        --            ##STR96##           I        -- Cl            ##STR97##           I        Cl            ##STR98##           I        Cl            ##STR99##           I        Cl     ______________________________________

                  TABLE 4     ______________________________________     Structural type:      ##STR100##     No.     X             Structure                                    Salt form     ______________________________________     69.     N(C.sub.2 H.sub.5).sub.2                           I     ______________________________________

                  TABLE 5     ______________________________________     Structural type:      ##STR101##     No     X                 Structure                                       Salt form     ______________________________________     70.    NHCH.sub.3        I     71.    NHC.sub.2 H.sub.5 I             ##STR102##       I             ##STR103##       I     74.    N(CH.sub.3)C.sub.2 H.sub.5                              I     ______________________________________

                  TABLE 6     ______________________________________     Structural type:      ##STR104##     No.     X                   Structure     ______________________________________              ##STR105##         I              ##STR106##         I     77.     N(CH.sub.3)C.sub.2 H.sub.5                                 I              ##STR107##         II              ##STR108##         I     ______________________________________

                  TABLE 7     ______________________________________     Structural type:      ##STR109##     No.  X                     Structure                                         Salt form     ______________________________________     82.  OC.sub.2 H.sub.5      III      --           ##STR110##           II       --           ##STR111##           II       --           ##STR112##           III      --           ##STR113##           I        --     87.  NHCH.sub.2CH(CH.sub.3).sub.2                                II       --     88.  NH(CH.sub.2).sub.3N(CH.sub.3).sub.2                                II       --           ##STR114##           II       --     ______________________________________

                  TABLE 8     ______________________________________     Structural type:      ##STR115##     No.    X                 Structure                                       Salt form     ______________________________________     90.    OC.sub.2 H.sub.5             ##STR116##       II       --     ______________________________________

                                      TABLE 9     __________________________________________________________________________      ##STR117##     Structural type I      ##STR118##     Structural type II     Compound           X                    Structural type                                       Mp °C.!     __________________________________________________________________________            ##STR119##          I       56-64     B            ##STR120##          I      176-184     (Salt form)     C            ##STR121##          I      166-168     D            ##STR122##          II     102-104     E (Cl)            ##STR123##          I      187     F (-)            ##STR124##          I       94-96     G (Cl)            ##STR125##          II     139-142     H (-)            ##STR126##          II     133-135     J (-)            ##STR127##          II     143-145     K (-)            ##STR128##          II      96-98     L (-)            ##STR129##          II     118-120     M (-)            ##STR130##          II     112-114     N (Cl)            ##STR131##          I       95-99     O (-)            ##STR132##          I      114-116     P (-)            ##STR133##          I       66-73     Q (Cl)            ##STR134##          II     205-209     __________________________________________________________________________

                                      TABLE 10     __________________________________________________________________________     Structural type:      ##STR135##     No.        X                      Structure                                    Salt form                                         Mp °C.!     __________________________________________________________________________     101         ##STR136##            I    Oxalate                                         138-140     102         ##STR137##            I    Oxalate                                         153-155     103         ##STR138##            I    Oxalate                                         117-123     104         ##STR139##            I    Oxalate                                         154-155     105         ##STR140##            I    Oxalate                                         127-129     106         ##STR141##            I    Oxalate                                         132-135     107         ##STR142##            I    Oxalate                                         145-147     108         ##STR143##            I    Base 118-120     109         ##STR144##            II   Base 118-120     110         ##STR145##            I    Oxalate                                         153-156     111        N (CH.sub.2).sub.5 CH.sub.3 !.sub.2                               I    Oxalate                                          81-84     112        N(CH.sub.2 CH.sub.2 CH.sub.3).sub.2                               I    Oxalate                                          57-59     113        N (CH.sub.2).sub.4 CH.sub.3 !.sub.2                               I    Oxalate                                         . . .     114         ##STR146##            I    Oxalate                                         125-126     115         ##STR147##            I    Base 103-105     116        N (CH.sub.2).sub.3 CH.sub.3 !.sub.2                               I    Oxalate                                         119-121     117        N CH.sub.2 CH(CH.sub.3).sub.2 !.sub.2                               I    Oxalate                                         130-131     118         ##STR148##            I    Oxalate                                         176-178     119         ##STR149##            I    Oxalate                                         160-162 (Decomp)     120         ##STR150##            I    Oxalate                                         170-172     121         ##STR151##            I    Oxalate                                         158-160 (Decomp)     122         ##STR152##            I    Oxalate                                         166-168     123        NH(CH.sub.2).sub.9CH.sub.3                               I    Oxalate                                          99-101     124         ##STR153##            I    Oxalate                                         127-143     125         ##STR154##            I    Oxalate                                         144-149     126         ##STR155##            I    Oxalate                                         137-139     127         ##STR156##            I    HCl  185-187     128        N(CH.sub.2 CN).sub.2   I    Base     129         ##STR157##            II   Base     130         ##STR158##            II   HCl     131         ##STR159##            II   HCl     132        NHCH.sub.2 -Adam(1)    II   HCl     133         ##STR160##            I    Oxalate     134         ##STR161##            I    Base     135         ##STR162##            I    HCl     136         ##STR163##            I    HCl     137         ##STR164##            I    (Oxalate).sub.2     138         ##STR165##            I    HCl     139         ##STR166##            I    HCl     140         ##STR167##            I    Base     141        OCH.sub.2C(CH.sub.3).sub.3                               II   Base     142         ##STR168##            II   Base     143        OCH.sub.2 CH.sub.2 -Adam(1)                               II   HCl     144        OCH.sub.2CH.sub.2C(CH.sub.3).sub.3                               II   HCl     145        OCH.sub.2 -Adam(1)     II   Base     146         ##STR169##            II   HCl     147         ##STR170##            II   HCl     148         ##STR171##            I    Oxalate                                         118-120     149         ##STR172##            I    HCl  195-197     150         ##STR173##            I    HCl  140-143     151         ##STR174##            I    HCl  193-195     152         ##STR175##            I    HCl  155-158     153         ##STR176##            I    HCl  168-170     154         ##STR177##            I    HCl  172-174     __________________________________________________________________________      Adam = Adamantyl

                                      TABLE 11     __________________________________________________________________________      ##STR178##     R.sub.1   X                  Salt form     __________________________________________________________________________      ##STR179##                ##STR180##        HCl  Mp. 116-127° C.      ##STR181##                ##STR182##        BS   Rf: 0.39 (ethyl acetate)     __________________________________________________________________________

                  TABLE 12     ______________________________________      ##STR183##     No.     R          Structure  Salt form                                           Mp °C.!     ______________________________________     1       4-CH.sub.3 I          Oxalate     2       4-F        I          Oxalate     3       3-Cl       I          Oxalate     4       4-Br       I          Oxalate     5       2-OCH.sub.3                        I          Oxalate     6       3-OCH.sub.3                        I          Oxalate     7       4-OCH.sub.3                        I          Oxalate     8       3,4-di-OCH.sub.3                        I          Oxalate     9       3,4,5-tri-OCH.sub.3                        I          Oxalate     10      4-N.sub.3  I          Oxalate     11      3-I,4-N.sub.3                        I          Oxalate     12      2-CH.sub.3 I          Oxalate     13      2-Br       I          Base     14      2,4-di-Cl  I          Oxalate     15      3-F        I          Oxalate     16      2-Cl       I          Base     ______________________________________

                                      TABLE 13     __________________________________________________________________________      ##STR184##     No.        NR.sup.3 R.sup.4     Salt form                                  Structure                                       Mp (°C.)                                            % Inhib.     __________________________________________________________________________      ##STR185##             1.5 Fu                                  I    177-178                                            40.44     "                       BS   II   158-159                                            0.0      ##STR186##             BS   II   165-167                                            50.64      ##STR187##             MS   I    132-133                                            67.73      ##STR188##             BS   II   84-55                                            65.50      ##STR189##             MS   I    151-153                                            80.89     "                       BS   II   110-111                                            39.47      ##STR190##             MS   I    118-122                                            24.03      ##STR191##             MS   I    150-154                                            43.49      ##STR192##             Cl   I    144-147                                            15.26      ##STR193##             BS   II   131-133                                            26.54      ##STR194##             Cl   I    114-115                                            0.0      ##STR195##             BS   II   144-147                                            67.75     __________________________________________________________________________

                                      TABLE 14     __________________________________________________________________________      ##STR196##     No.        NR.sup.3 R.sup.4        Salt form                                     Structure                                          Mp (°C.)                                                % Inhib.     __________________________________________________________________________      ##STR197##                Cl   I    amorphous.sup.x)                                                79.17      ##STR198##                Cl   I    amorphous.sup.x)                                                73.55      ##STR199##                Cl   I    amorphous.sup.x)                                                60.82      ##STR200##                Cl   I + II                                          amorphous.sup.x)                                                82.76      ##STR201##                Cl   I    amorphous.sup.x)                                                50.90      ##STR202##                Ox   I    amorphous.sup.x)                                                56.20      ##STR203##                Ox   I    142-144      ##STR204##                BS   I    105-106                                                22.39      ##STR205##                Cl.sub.2                                     I    208-210      ##STR206##                MS   I    amorphous.sup.x)                                                79.60      ##STR207##                Cl   I    amorphous.sup.x)                                                36.71      ##STR208##                Cl   I    amorphous.sup.x)                                                30.87     __________________________________________________________________________      .sup.x) The structure of the substance is characterised by NMR      spectroscopy

                                      TABLE 15     __________________________________________________________________________      ##STR209##     No.        NR.sup.3 R.sup.4  Salt form                               Structure                                    Mp (°C.)                                         % Inhib.     __________________________________________________________________________      ##STR210##          Cl   I    134-136      ##STR211##          Cl   I    166-168      ##STR212##          Cl   I    142-144      ##STR213##          BS   I    119  84.43     __________________________________________________________________________

                                      TABLE 16     __________________________________________________________________________      ##STR214##        Salt     No.        form           R.sup.a     R.sup.b    NR.sup.3 R.sup.4    Mp.  %                                                              IC.sub.50     __________________________________________________________________________     MS    HO          CH.sub.3 O                                   ##STR215##              87 3.47 ×                                                              10.sup.-6     MS    CH.sub.3 O  HO         "                        0  --     MS    CH.sub.3 O  CH.sub.3 O                                   ##STR216##     OX    CH.sub.3 O  H                                   ##STR217##         60-70                                                           30.41     MS            ##STR218## H          "                   182-185                                                           14.65     OX    HO          H          "                   120-130     OX    CH.sub.3 O  CH.sub.3   "                   143  10.47     OX            ##STR219## CH.sub.3   "                   156-157                                                           49.03     OX    HO          CH.sub.3   "                   80-95     OX    CH.sub.3 O  Cl         "                   143-145                                                           44.65     OX            ##STR220## Cl         "                   130-132                                                           32.90     OX    HO          Cl         "                   182-183     MS            ##STR221## CH.sub.3 O "                        31.8     MS    CH.sub.3 O                        ##STR222##                                  "                        22.9     OX    CH.sub.3 O  CH.sub.3                                   ##STR223##         145-151                                                           58.3     OX    CH.sub.3 O  H          "                   115-125     OX    CH.sub.3 O  Cl         "                   131-135     OX            ##STR224## Cl         "                   135-142     OX            ##STR225## CH.sub.3   "                   162-164     OX            ##STR226## H          "                   145-147     OX    HO          Cl         "                   187-190     OX    HO          CH.sub.3   "                   193-200     OX    HO          H          "                   154-157     __________________________________________________________________________

                                      TABLE 17     __________________________________________________________________________      ##STR227##     No.        NR.sup.3 R.sup.4       Salt form                                    Mp(°C.)                                          % Hem.                                              IC.sub.50     __________________________________________________________________________      ##STR228##               OX   amorphous*                                          68.2      ##STR229##               Cl   amorphous*                                          36.95      ##STR230##               Cl         69.7      ##STR231##               Cl   amorphous*      ##STR232##               BS   amorphous*      ##STR233##               Cl   amorphous*     HNCH.sub.2 -1-Adam.       Cl   amorphous*                                          65.1      ##STR234##               BS   amorphous*      ##STR235##               Cl   amorphous*                                          12.82      ##STR236##               BS   amorphous*                                          59      ##STR237##               Cl   amorphous*                                          72.18      ##STR238##               Cl   amorphous*      ##STR239##               OX-2 amorphous*                                          41.06      ##STR240##               BS   amorphous*                                          7.77      ##STR241##               BS   amorphous*                                          45.26      ##STR242##               OX   amorphous*                                          33.06      ##STR243##               Cl   199-201                                          81.60      ##STR244##               Cl   amorphous*                                          52.51      ##STR245##               Cl   amorphous*                                          80.97                                              7.57 × 10.sup.-6     __________________________________________________________________________      *The structure of the substance is characterised by NMR spectroscopy

                  TABLE 18     ______________________________________      ##STR246##                          Salt     No.  R.sup.1         form   Mp.     % Hem.                                               IC.sub.50     ______________________________________      ##STR247##      OX     amorphous*                                       41.8      ##STR248##      OX     amorphous*                                       24.3      ##STR249##      OX     amorphous*                                       73.7      ##STR250##      OX     amorphous*                                       53.1      ##STR251##      OX     amorphous*      ##STR252##      OX     amorphous*                                       68.22      ##STR253##      OX     amorphous*      ##STR254##      BS     amorphous*      ##STR255##      OX     amorphous*      ##STR256##      BS     amorphous*      ##STR257##      OX     amorphous*      ##STR258##      OX     amorphous*                                       30.79      ##STR259##      OX     amorphous*      ##STR260##      OX     amorphous*      ##STR261##      OX     amorphous*                                       54.30      ##STR262##      OX     amorphous*                                       61.3      ##STR263##      Cl     amorphous*                                       5.81      ##STR264##      BS     amorphous*      ##STR265##             amorphous*                                       56.0      ##STR266##      OX     amorphous*      ##STR267##      OX     amorphous*     ______________________________________      *: The structure of the substance is characterised by NMR spectroscopy

                  TABLE 19     ______________________________________      ##STR268##     ______________________________________      *: The structure of the substance is characterised by NMR spectroscopy

                                      TABLE 20     __________________________________________________________________________      ##STR269##     No.        R.sup.a R.sup.b                     NR.sup.3 R.sup.4  Salt form                                            Mp.  % H                                                    IC.sub.50     __________________________________________________________________________     HO         CH.sub.3 O                      ##STR270##       MS   163-169                                                 86.70                                                    3.47 × 10.sup.-6     PhCH.sub.2O                CH.sub.3 O                     "                 MS   amorph*                                                 31.78     PhCH.sub.2 O                CH.sub.3 O                     NHCH.sub.2 CH.sub.2C(CH.sub.3).sub.3                                       OX   amorph*                                                 61.85     PhCH.sub.2O                Cl                      ##STR271##       OX   130-132                                                 32.9     CH.sub.2 O Cl   "                 OX   143-145                                                 44.65     PhCH.sub.2 O                CH.sub.3                     "                 OX   156-157                                                 49.03     __________________________________________________________________________      *The structure of the substance is characterised by NMR spectroscopy

The present invention further relates to the use of the above-mentioned compounds for the production of agents for treating chronic inflammatory processes, ulcerative colitis and Crohn's disease and for producing agents having an antiproliferative effect. The activity of the compounds can be explained by their inhibition of the unselective cation channels (UCC).

The pathophysiology of chronic bronchial asthma is based on inflammatory processes which are mediated by the activation of inflammatory cells. (BARNES, 1987; SEIFERT and SCHULTZ, 1991).

The receptor-regulated activation of inflammatory cells (e.g. neutrophilic granulocytes and mast cells or the permanent cell lines HL-60 cells or sensitised RBL cells, i.e. those charged with gammaglobulin E) is inhibited, irrespective of the nature of the stimulating agonists (e.g. endothelin, PAF, leukotrienes, chemotactical peptide fMLP or antigen against sensitised mast cells) by blockers of unselective cation channels (UCC) (RINK, 1990). Through these channels extracellular calcium, which is responsible for the persistence of receptor-mediated cell activations, enters the cells (PUTNEY, 1990). If this supply of calcium is interrupted this results in a blockade of the activation of inflammatory cells.

Conventional calcium antagonists of the dihydropyridine or phenylalkylamine type do not inhibit either UCCs or inflammatory processes (WELLS et al., 1986).

As a measurement of the cell activation or as a measurement of the inhibition thereof by UCC blockers, the kinetics of the cytoplasmic calcium ion concentration in fura-2-charged cells is quantified fluorometrically using the method described by GRYNKIEWICZ et al. (1985). This procedure has proved a reliable screening method, within the scope of the invention, for detecting UCC blockers.

So-called functional THAPSIGARGIN inhibition has proved suitable for the specific characterisation of blockers of the unselective cation channels. THAPSIGARGIN is a tumour promoter described by THASTRUP et al. (Proc. Natl. Acad. Sci. (USA), 87, 2466-2470, 1990) which selectively and irreversibly inhibits the Ca²⁺ -ATPase of intracellular IP₃ -sensitive Ca²⁺ -stores. Consequently the Ca²⁺ -stores are rapidly depleted. As described by J. PUTNEY (Calcium, 11, 611-624, 1990) the depletion of these stores constitutes the physiological stimulation for opening up unselective cation channels in the cell membrane. The result of this is a massive influx of Na⁺ and Ca²⁺ into the cell. Because of these properties, Thapsigargin is suitable as an indirect stimulator for agonist- and IP₃ -independent opening up of the unselective cation channels.

Within the scope of the present invention the Thapsigargin stimulation of unselective cation channels has been carried out successfully on HL 60 cells (human leukaemia cells), on hippocampal and cortical neurone cells and on RBL-cells (rat basophilic lymphoma cells) and in this way the existence of these channels in particular cell lines was demonstrated.

The cytoplasmic Ca²⁺ concentration ( Ca²⁺ !_(i)) plays an important part in the cell proliferation and in tumour growth (for a summary see L. R. ZACHARSKI, Journal of Medicine 19: 145-177, 1988). In particular, the Ca²⁺ -influx into the cell stimulated by receptor activation with consecutive inositoltriphosphate-(IP₃ -)-mediation would appear to be of crucial importance for oncogenic cell proliferation (U. KIKKAWA and Y. NISHIZUKA, Ann. REV. CELL. BIOL. 2: 149-178, 1986). This mechanism also plays a part in the formation of metastases and in "Multi-Drug Resistance". (For a summary see the above-mentioned publication by L. R. ZACHARSKI, J. MED. 19: 145-177, 1980).

This hypothesis is supported by the fact that Thapsigargin, as an indirect stimulator of the unselective cation channels (UCC) not only leads to a Ca²⁺ -overload in the cell but is also a highly effective tumour promoter. (V. THASTRUP et al. Proceedings of the NATL. Acad. Sci: (USA) 87: 2466-2470, 1990).

The blockade of the Ca²⁺ -influx by the UCC leads to normalisation of the intracellular Ca-ion concentration and hence to inhibition of tumour growth etc.

Conventional calcium antagonists do not inhibit these UCC. It has been found, surprisingly, that the compounds according to this invention inhibit the influx of calcium into the cell through the UCC.

As shown by S. H. MURCH et al. (Lancet 339: 381-385, 15. Feb. 1992) endothelin I plays an important pathophysiological role in inflammatory intestinal diseases such as ulcerative colitis and Crohn's disease. Using immunohistochemical methods it has been shown that patients with Crohn's disease in the region of the submucosa and patients with ulcerative colitis in the region of the lamina propria of the epithelium of the large intestine show significantly and greatly increased concentrations of endothelin I compared with healthy normal people. It is assumed that the local secretion of endothelin causes massive vasospasms with consecutive disseminated ischaemia with microinfarcts which are regarded as the actual cause of the above diseases. The vasospasmogenic effectiveness of endothelin is explained by a Ca²⁺ -overload of vascular myocytes. Endothelin primarily triggers an IP₃ -mediated intracellular release of Ca²⁺ which is followed by a massive transmembranal Ca²⁺ -entry through dihydropyridine-insensitive channels. (M. S. Simonson et al. Clin. Invest. Med. 14: 499-507, 1991; T. Masakai, J. Cardiovasc. Pharmacol. 13:Suppl. 5, S1-S4, 1989; D. W. Hay, R. J. Pharmacol. 100: 383-392, 1990). These channels are unselective cation channels which have also been briefly described as existing in cells of the large intestine mucosa. (Chr. Siemer and H. Gogelein, Europ. J. Physiol. 420: 319-328, 1992).

The endothelin-stimulated activation of fura-2-charged human leukaemia cells (HL 60 cells) has proved a suitable screening model for detecting functional endothelin antagonists. In conformity with G. GRYNKIEWICZ et al. (J. Biol. Chem. 260:3440-3450, 1985) the intracellular Ca²⁺ -concentration in the cytoplasm of HL 60 cells (suspensions) can be monitored by spectrofluorometry and quantified as a measurement of cell activation by endothelin. The stimulation was effected by adding 0.1 μM endothelin and could be inhibited in a dosage-dependent manner by means of the substances according to the invention.

The functional endothelin antagonism of the substances according to the invention is mediated through a blockade of the unselective cation channels. Consequently, detection of a functional Thapsigargin-antagonism on RBL-hm1 cells is also a suitable screening method for functional endothelin antagonists.

Carrying out the investigation:

For screening purposes, fura-2-charged adhesive RBL-hm 1 cells are stimulated with 0.1 μM Thapsigargin in a Ca²⁺ -free incubation medium. After 4 minutes, extracellular Ca²⁺ is restored to a concentration of 1.5 mM and, using the fura-2-fluorescence, the excessive increase in the cytoplasmic Ca²⁺ -concentration caused by a massive transmembranal Ca²⁺ -entry through unselective cation channels is recorded.

This entry is to be inhibited solely by unselective cation channel blockers in a dosage-dependent manner. Neither conventional calcium antagonists nor specific blockers of agonists which stimulate the IP₃ -turnover are able to inhibit the transmembranal Ca²⁺ -entry triggered indirectly by Thapsigargin. The compounds of the present invention are distinguished by their inhibition of UCC.

The fluorometric calcium measurement in the cytoplasm of individual adhering RBL-hm1 cells is carried out analogously to the method described by KUDO and OGURA (1986) for neuronal cells. An AXIOVERT 35 fluorescence microscope made by ZEISS is used in conjunction with an imaging system made by HAMAMATSU, consisting of the ICMS-image processing system, residual light camera with control unit and image intensifier DVS 3000.

The kinetics of the cytoplasmic Ca²⁺ -concentration is recorded continuously as a concentration/time curve after the cell activation stimulated by Thapsigargin (0.1 μM). The curves of two activated cell cultures are compared in the presence and absence of 10 μM test substance. The area under these curves (area under the curve=AUC) is integrated and recorded as a measurement of cell activation. The inhibitory potency of the UCC-blockers tested is determined using the following equation: ##EQU1## %H=the percentage inhibition of the calcium entry through unselective cation channels which is stimulated and inhibited by 10 μM of test substance.

AUC_(inh) =area under the curve recorded in the presence of the stimulant plus 10 μM inhibitory test substance.

AUC_(control) =area under the curve which is recorded only after the addition of the stimulant.

LITERATURE RELATING TO THE ABOVE EXPLANATIONS

BARNES P. J., I. W. RODGER and N. C. THOMSON Pathogenesis of asthma, in "ASTHMA, basic mechanisms and clinical management"ED by P. J. BARNES; ACADEMIC PRESS, LONDON, 1988

GRYNKIEWICZ G., M. POENIE and R. Y. TSIEN A new generation of Ca²⁺ -indicators with greatly improved fluorescence properties J. BIOL. CHEM. 260: 3440-3450, 1985

HIDE, M. and M. A. BEAVEN Calcium influx in a rat mast cell (RBL-2H3) line J. BIOL. CHEM. 266 15221-15229, 1991

KUDO, Y. and A. OGURA Glutamate-induced increase in intracellular Ca²⁺ -concentration in isolated hippocampal neurones BR. J. PHARMACOL. 89: 191-198, 1986

PUTNEY, J. W., jr. Capacitative Calcium entry revised CELL CALCIUM 11: 611-624, 1990

RINK, T. J. Receptor-mediated calcium entry FEBS LETT. 268: 381-385, 1990

SEIFERT, R. and G. SCHULTZ The superoxide forming NADPH oxidase of phagocytes: An enzyme system regulated by multiple mechanism REV. PHYSIOL. BIOCHEM. PHARMACOL., Vol. 117, SPRINGER VERL., 1991

WELLS, E., C. G. JACKSON, S. T. HARPER, J. MANN and R. P. EAOY

Characterization of primate bronchoalveolar mast cells II, inhibition of histamine, LTC₄ and PGF₂α release from primate bronchoalveolar mast cells and a comparison with rat peritoneal mast cells

J. IMMUNOL. 137: 3941-3945, 1986.

Results of measurement:

The percentage inhibition of UCC after Thapsigargin stimulation (0.1 μM Thapsigargin) in RBL-hm 1 cells is given. The uniform concentration of the test substances is 10⁻⁵ mol).

                  TABLE 21     ______________________________________     (Data relating to compounds of Tables 1-10)     RBL - hm 1 cells - Thapsigargin (0.1 μM) stimulation     Compound     No.            % Inhibition     ______________________________________     3              F 41.91     4              F 66.53     5              F 80.16     9              F 67.59     14             F 80.16     17             F 52.53     18             F 57.53     20             F 57.38     33             60.55     36             F 52.19     37             F 75.77     38a (oxalate)  52.64     39             48.62     40             64.99     41 (Structure I)                    F 57.22     42             84.83     43             F 48.69     46             68.06     48             61.34     83             66.76     89             53.38     B              72.35     C              63.45     E              F 81.89     F              48.01     G              57.52     H              57.06     K              76.31     L              53.14     O              63.32     Q              64.28     101            59.62     102            F 80.77     104            54.67     105            45.09     106            F 65.69     108            44.45     109            74.18     115            75.53     132            F 65.14     133            68.15     ______________________________________

The activity data (%inhibition and IC₅₀) for other compounds are contained in Tables 13-20.

The functional antiinflammatory effectiveness can be demonstrated by means of the following test:

Individual RBL-2H3-cells (a tumour cell line related to the mast cells) adhering to glass slides are used.

The cultivation and attachment of the RBL-2H3-cells are carried out by the method described by HIDE and BEAVEN (1991). In order to sensitise the adhesive RBL-2H3-cells the cells are incubated for 2 hours at ambient temperature with a 1:2000 diluted commercial gammaglobulin E-solution against a dinitrophenol-bovine serum albumin complex (DNP-BSA-antigen). The cells are then washed. By the addition of 0.1 ml of DNP-BSA-solution (10 μg/ml) there is a massive immunological cell activation which is mediated by a cytoplasmic Ca²⁺ -overload. The fluorometric calcium measurement in the cytoplasm of individual adhering RBL-2H3-cells is carried out analogously to the method described by KUDO and OGURA (1986) for neuronal cells, which is also explained hereinbefore in this specification.

The comparison used in these investigations is (-10 μM) chromoglycate which brings about an approximately 50% inhibition of the antigen-induced cell activation.

In this test the above-mentioned compounds demonstrate %H values which are comparable with the values specified hereinbefore.

Tests on microcultures of various human tumour cell lines using the tetrazolium assay in order to determine the antiproliferative effect of the substances according to the invention surprisingly showed that the compound tested was 5 to 100 times more potent than the comparison substance Verapamil.

The antiproliferative effectiveness of the test substances was determined by means of the MTT test described by MOSMANN (J. IMMUNOL. METH. 65: 55-63, 1983), DENIZOT et al. (J. IMMUNOL. METH. 89: 271-277, 1986) and J. ELIASON et al. (INT. J. CANCER 46: 113-117, 1990). (MTT= 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide! produced by CHEMICON Inc. El Segundo, Calif., USA). This indicator is metabolised only by living cells with intact mitochondria into a blue formazane product. The following human tumour cell lines were used in our test: A 549 (adenocarcinoma of the lung), A 431 (epidermal carcinoma of the vulva), PC 3 (adenocarcinoma of the prostate), SK BR 3 (adenocarcinoma of the breast), HT 29 (CX1 1) (adenocarcinoma of the colon) and K 562 (chronic myeloid leukaemia cell). The test was carried out on microtitre plates. Each well contained 100 μl of a cell suspension (0.2×10⁶ cells per ml). The incubation medium used was RPMI 1640 with 10% heat-inactivated foetal calves' serum and 50 μg/ml of gentamycin. The cell suspensions were incubated for 0, 24, 48 or 72 hours in air with a humidity at saturation point in a CO₂ (5%)/air (95%) mixture at 37° C., incubated in the presence and absence of variable concentrations of antiproliferative test substances. The test substances were dissolved in DMSO (final dilution: 0.1%). Then 10 μl of MTT-solution (3 mg/ml) were added, followed after 3 hours by 100 μl of an isopropanol solution containing 0.08N HCl. After a further hour, the light absorption at 570 nm (comparative wavelength 630 nm) was determined in a microplate reader. The light absorption is directly proportional to the number of living cells. The half-maximum inhibitory concentrations of the substances tested were 1 μg/ml.

The vasospasmolytic effectiveness of the above-mentioned functional endothelin and Thapsigargin antagonists were confirmed on an isolated blood vessel preparation: coronary perfusion was continuously quantified, on retrogressively perfused, spontaneously beating LANGENDORFF hearts taken from rats, by means of electromagnetic flow measurement (apparatus supplied by Hugo Sachs Elektronik, MARCH). This measuring apparatus could be used to record the extent, duration and pattern of vascular spasms with a high degree of accuracy. If perfusion is carried out with 100 nM endothelin concentration, the coronary perfusion flow is reduced from 11 to 5 ml/min. The restriction in perfusion can be reversed by means of the substances according to the invention. The potencies of the compounds according to the invention with regard to Thapsigargin inhibition on fura-2-charged RBL-hm1-cells or the effectiveness of endothelin-inhibition on fura-2-charged HL 60 cells correlates clearly with the vasospasmolytic effectiveness of the test substances detected on the Langendorff preparation. It can be concluded from this that, underlying the vasospasmolytic endothelin antagonism of the substances tested, there is a blockade of the unselective cation channels.

EXAMPLES OF PHARMACEUTICAL PREPARATIONS

    ______________________________________     a) Coated tablets     ______________________________________     1 tablet core contains:     Active substance of general formula I                              30.0   mg     Lactose                  100.0  mg     Corn starch              75.0   mg     Gelatine                 3.0    mg     Magnesium stearate       2.0    mg                              210.0  mg     ______________________________________

Preparation

The active substance mixed with lactose and corn starch is granulated with a 10% aqueous gelatine solution through a 1 mm mesh screen, dried at 40° C. and rubbed through a screen once more. The granules thus obtained are mixed with magnesium stearate and compressed. The cores produced in this way are coated in the usual manner with a coating consisting of an aqueous suspension of sugar, titanium dioxide, talc and gum arabic. The finished coated tablets are polished with beeswax.

    ______________________________________     b) Tablets     ______________________________________     Active substance of general formula I                              30.0   mg     Lactose                  100.0  mg     Corn starch              70.0   mg     Soluble starch           7.0    mg     Magnesium stearate       3.0    mg                              210.0  mg     ______________________________________

Preparation

The active substance and magnesium stearate are granulated with an aqueous solution of the soluble starch, the granules are dried and intimately mixed with lactose and corn starch. The mixture is then compressed into tablets weighing 210 mg.

    ______________________________________     c) Capsules     ______________________________________     Active substance according to claim 1                              20.0   mg     Lactose                  230.0  mg     Corn starch              40.0   mg     Talc                     10.0   mg                              300.0  mg     ______________________________________

Preparation

The active substance, lactose and corn starch are first combined in a mixer and then in a grinding machine. The mixture is returned to the mixer, thoroughly combined with the talc and mechanically packed into hard gelatine capsules.

    ______________________________________     d) Tablets     ______________________________________     Active substance according to the invention                               40.0   mg     Lactose                   100.0  mg     Corn Starch               50.0   mg     Colloidal silica          2.0    mg     Magnesium stearate        3.0    mg                          Total    200.0  mg     ______________________________________

Preparation

The active substance is mixed with some of the excipients and granulated with a solution of the soluble starch in water. After the granules have been dried the remaining excipients are added and the mixture is compressed to form tablets.

    ______________________________________     e) Coated tablets     ______________________________________     Active substance according to the invention                               20.0   mg     Lactose                   100.0  mg     Corn starch               65.0   mg     Colloidal silica          2.0    mg     Soluble starch            5.0    mg     Magnesium stearate        3.0    mg                          Total    195.0  mg     ______________________________________

Preparation

The active substance and excipients are compressed to form tablet cores as described in Example 1 which are then coated with sugar, talc and gum arabic in the usual way.

    ______________________________________     f) Suppositories     ______________________________________     Active substance according to the invention                               50.0   mg     Lactose                   250.0  mg     Suppository mass q.s. ad  1.7    g     ______________________________________

Preparation

The active substance and lactose are mixed together and the mixture is uniformly suspended in the molten suppository mass. The suspensions are poured into cooled moulds to form suppositories weighing 1.7 g.

    ______________________________________     g) Ampoules     ______________________________________     Active substance according to the invention                               20.0   mg     Sodium chloride           5.0    mg     Twice distilled water q.s. ad                               2.0    ml     ______________________________________

Preparation

The active substance and sodium chloride are dissolved in twice distilled water and the solution is transferred under sterile conditions into ampoules.

    ______________________________________     h) Ampoules     ______________________________________     Active substance according to the invention                               10.0   mg     Sodium chloride           7.0    mg     Twice distilled water q.s. ad                               1.0    mg     ______________________________________

    ______________________________________     i) Drops     ______________________________________     Active substance according to the invention                              0.70    g     Methyl p-hydroxybenzoate 0.07    g     Propyl p-hydroxybenzoate 0.03    g     Demineralised water q.s. ad                              100.00  ml     ______________________________________

Preparation

The active substance and preservatives are dissolved in demineralised water, the solution is filtered and transferred into vials holding 100 ml. 

What is claimed is:
 1. A compound of formula ##STR272## wherein A is benzo m is 2 or 3R² is hydroxy, (C₁₋₄)alkoxy, benzyloxy, halogen, (C₁₋₄)alkyl, methanesulphonyloxy or methanesulphonamido or two adjacent R² substituents together are --O--CH₂ --O-- or --O--CH₂ --CH₂ --O-- R¹ is ##STR273## Wherein R is C₁₋₄ -alkyl, hydroxy, --N₃, halogen, CF₃, C₁₋₄ -alkoxy or --COH and u is 1, 2 or 3 R³ and R⁴ independently of each other are (a) hydrogen (b) branched or unbranched C₃₋₆ -alkenyl, (c) branched or unbranched C₃₋₆ -alkynyl, or (d) branched or unbranched C₁₋₁₂ -alkyl, whilst the alkyl may be substituted by hydroxy (C₁₋₄) alkoxy di(C₁₋₄)alkylamino furyl pyridyl pyrrolidinyl, N-methyl pyrrolidinyl, morpholino indolyl nitrilo thienyl adamantyl cyclohexyl phenoxy naphthyloxy or phenyl, whilst this phenyl or the phenyl contained in the phenoxy group may be mono-, di- or tri-substituted by hydroxy, (C₁₋₄)alkoxy, benzyloxy, halogen, CF₃, N₃, (C₁₋₄)alkyl, adamantyl, --SO₂ NH₂, --NHCOCH₃, --NHSO₂ CH₃ or CH₃ SO₂ O-- or by the bridge --O--CH₂ --O--; or R³ is hydrogen and R⁴ is cyclohexyl, phenyl, fluorophenyl, pyridyl or N-benzylpiperidyl; or R³ and R⁴ together with the nitrogen atom to which they are bound represent pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, the group ##STR274## or piperazinyl, whilst the piperazinyl ring may optionally be N-substituted by methyl, unsubstituted phenyl, mono- or di- (C₁₋₄) alkoxyphenyl, pyrimidinyl, phenyl (C₁₋₄)alkyl or ##STR275## or a salt thereof with a physiologically acceptable acid, base or complexing agent.
 2. The compound as recited in claim 1 wherein m is 2, and the two R² are in positions 6 and
 7. 3. The compound as recited in claim 1 wherein NR³ R⁴ is ##STR276##
 4. The compound as recited in claim 1 wherein R is methoxy, methyl, F or N₃.
 5. A method for treating chronic inflammatory processes, ulcerative colitis and Crohn's disease in a warm-blooded animal, which comprises administering to said animal a therapeutically effective amount of the compound of claim
 1. 6. A method for treating diseases in a warm-blooded animal that respond to an agent with anti-proliferative activity which comprises administering to said animal a therapeutically effective amount of the compound of claim
 1. 